PDF(Pubmed)
Abstract：
This study presents the chemical synthesis, purification, and characterization of a novel non-natural synthetic amino acid. The compound was synthesized in solution, purified, and characterized using NMR spectroscopy, polarimetry, and melting point determination. Dynamic Light Scattering (DLS) analysis demonstrated its ability to form aggregates with an average size of 391 nm, extending to the low micrometric size range. Furthermore, cellular biological assays revealed its ability to enhance fibroblast cell growth, highlighting its potential for tissue regenerative applications. Circular dichroism (CD) spectroscopy showed the ability of the synthetic amino acid to bind serum albumins (using bovine serum albumin (BSA) as a model), and CD deconvolution provided insights into the changes in the secondary structures of BSA upon interaction with the amino acid ligand. Additionally, molecular docking using HDOCK software elucidated the most likely binding mode of the ligand inside the BSA structure. We also performed in silico oligomerization of the synthetic compound in order to obtain a model of aggregate to investigate computationally. In more detail, the dimer formation achieved by molecular self-docking showed two distinct poses, corresponding to the lowest and comparable energies, with one pose exhibiting a quasi-coplanar arrangement characterized by a close alignment of two aromatic rings from the synthetic amino acids within the dimer, suggesting the presence of π-π stacking interactions. In contrast, the second pose displayed a non-coplanar configuration, with the aromatic rings oriented in a staggered arrangement, indicating distinct modes of interaction. Both poses were further utilized in the self-docking procedure. Notably, iterative molecular docking of amino acid structures resulted in the formation of higher-order aggregates, with a model of a 512-mer aggregate obtained through self-docking procedures. This model of aggregate presented a cavity capable of hosting therapeutic cargoes and biomolecules, rendering it a potential scaffold for cell adhesion and growth in tissue regenerative applications. Overall, our findings highlight the potential of this synthetic amino acid for tissue regenerative therapeutics and provide valuable insights into its molecular interactions and aggregation behavior.
摘要：
本研究提出了化学合成，净化,并对一种新型非天然合成氨基酸进行了表征。该化合物是在溶液中合成的，纯化,并使用NMR光谱进行表征，偏振法,和熔点测定。动态光散射（DLS）分析表明其能够形成平均尺寸为391nm的聚集体，延伸到低微米尺寸范围。此外，细胞生物学分析显示其增强成纤维细胞生长的能力，突出了其组织再生应用的潜力。圆二色性（CD）光谱显示合成氨基酸结合血清白蛋白的能力（使用牛血清白蛋白（BSA）作为模型），和CD去卷积提供了与氨基酸配体相互作用时BSA二级结构变化的见解。此外,使用HDOCK软件的分子对接阐明了BSA结构内配体最可能的结合模式。我们还对合成化合物进行了硅低聚，以获得聚集体模型以进行计算研究。更详细地说,通过分子自对接实现的二聚体形成显示出两种不同的姿势，对应于最低和可比的能量，一个姿势表现出准共面排列，其特征是二聚体中合成氨基酸的两个芳环紧密对齐，表明存在π-π堆叠相互作用。相比之下,第二个姿势显示非共面的配置，芳香环以交错排列的方式排列，指示不同的交互模式。在自对接程序中进一步利用了这两种姿势。值得注意的是,氨基酸结构的迭代分子对接导致高阶聚集体的形成，使用通过自对接程序获得的512聚体聚集体的模型。这种聚集体模型呈现了一个能够容纳治疗性货物和生物分子的空腔，使其成为组织再生应用中细胞粘附和生长的潜在支架。总的来说,我们的发现强调了这种合成氨基酸在组织再生治疗中的潜力，并为其分子相互作用和聚集行为提供了有价值的见解。
